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Strain domestication and screening method based on micro-fluidic technology

A microfluidic technology and screening method technology, which can be used in microorganism-based methods, methods using microorganisms, biochemical equipment and methods, etc.

Pending Publication Date: 2021-05-18
LUOYANG TMAXTREE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The entire domestication process cannot be fully automated, and it is very easy to infect bacteria during the artificial passaging process. The passaging operation and growth monitoring of the strains will consume a lot of manpower, and the detection requires sampling operations. Due to the limitation of the sample volume, intensive detection cannot be carried out. The amount of data obtained is very limited

Method used

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  • Strain domestication and screening method based on micro-fluidic technology
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Escherichia coli domestication and screening

[0038] 1. Experimental materials and reagents

[0039] Material: Escherichia coli DH 5α

[0040] Reagents: LB solid medium: agar 20g, tryptone 10g, yeast extract 5g, NaCl 10g, distilled water to volume 1L, pH 7.0, sterilized at 121°C for 20 min.

[0041] LB liquid medium A: tryptone 10g, yeast extract 5g, NaCl 10g, distilled water to volume 1L, pH 7.0, sterilized at 121°C for 20 min.

[0042] LB liquid medium B: tryptone 10g, yeast extract 5g, NaCl 30g, distilled water to volume 1L, pH 7.0, sterilized at 121°C for 20 min.

[0043]2. Experimental equipment: automatic high-throughput microbial droplet culture system (Microbial Microdroplet Culturesystem, MMC).

[0044] 3. Experimental steps:

[0045] 1) Inoculate Escherichia coli DH 5α on LB solid medium and culture it at 37°C. After the cultivation, pick a single colony and inoculate it in 5mL LB liquid medium A, and culture it at 37°C with shaking at 200 r...

Embodiment 2

[0058] Example 2: Saccharomyces cerevisiae domestication and screening

[0059] Saccharomyces cerevisiae can convert sugars into ethanol, which is widely used in industry. However, the high concentration of ethanol produced in the production process will inhibit the growth of Saccharomyces cerevisiae. In order to obtain Saccharomyces cerevisiae that tolerates high concentrations of ethanol, it is necessary to domesticate Saccharomyces cerevisiae with ethanol resistance.

[0060] 1. Experimental materials and reagents

[0061] Material: Saccharomyces cerevisiae

[0062] Reagents: YPD solid medium: agar 20g, peptone 20g, yeast extract 10g, glucose 20g, distilled water to volume 1L, sterilized at 115°C for 15 min.

[0063] YPD liquid medium A: peptone 20g, yeast extract 10g, glucose 20g, distilled water to volume 1L, sterilized at 115°C for 15 min. Add ethanol to a volume fraction of 10%.

[0064] YPD liquid medium B: peptone 20g, yeast extract 10g, glucose 20g, distilled wat...

Embodiment 3

[0086] Example 3 Domestication and Screening of Lactobacillus plantarum

[0087] Lactobacillus plantarum is a strain widely used in the food industry, and the product lactic acid in the fermentation process can inhibit the growth of the strain. In order to relieve the inhibitory effect of this product, it is necessary to domesticate Lactobacillus plantarum for lactic acid resistance.

[0088] 1. Experimental materials and reagents

[0089] Material: Lactobacillus plantarum

[0090] Reagents: MRS solid medium: agar 20g, peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, glucose 20g, Tween 80 1mL, sodium acetate 5g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58 g, manganese sulfate 0.25g, distilled water to volume 1L, pH6.2~6.6, sterilized at 115℃ for 15min.

[0091] MRS liquid medium A: peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, glucose 20g, Tween 80 1mL, sodium acetate 5g, dipotassium hydrogen phos...

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Abstract

The invention relates to a strain domestication and screening method based on a micro-fluidic technology, and belongs to the application of the microfluidic technology in the field of biology. According to the invention, strain domestication and screening are carried out based on a microbial microdroplet culture mode; The microbial microdroplet culture mode is realized through high-throughput microbial droplet culture equipment, a microdroplet unit is formed in a micro-fluidic chip of the equipment, online continuous culture is performed, chemical factor culture media with different concentrations can be replaced in the process to realize strain domestication, and finally, by means of OD screening, the optimal microdroplet unit is picked out and extracted.

Description

technical field [0001] The invention belongs to the application of microfluidic technology in the field of microbial cultivation, and in particular relates to a method for acclimating and screening strains based on microfluidic technology. Background technique [0002] Strain domestication refers to the method of directional breeding of microorganisms by gradually adapting them to a certain condition through artificial measures. The usual method is to use the shake flask culture method to subculture the strains multiple times, increase the growth pressure (usually some chemical factors) during the passage of the strains, and finally obtain the strains that can tolerate high growth pressure. [0003] The entire domestication process cannot be fully automated, and it is very easy to infect bacteria during the artificial passaging process. The passaging operation and growth monitoring of the strains will consume a lot of manpower, and the detection requires sampling operations....

Claims

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Application Information

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IPC IPC(8): C12N1/36C12N1/20C12N1/18C12R1/19C12R1/865C12R1/25
CPCC12N1/36C12N1/20C12N1/18
Inventor 王立言郭肖杰张乐乐
Owner LUOYANG TMAXTREE BIOTECH CO LTD
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